Date of Award
12-2009
Degree Type
Thesis
Degree Name
Master of Science (MS)
Department
Chemistry
First Advisor
Dr. Jenny Yang - Chair
Second Advisor
Dr. Giovanni Gadda
Third Advisor
Dr. Zhi-Ren Liu
Abstract
Calcium plays important roles in both eukaryotic and prokaryotic cells. Its actions help to stabilize cell synthesis, growth and development. In this thesis, studies have been completed to determine effects of calcium and pH on bacterial cell growth and protein expression using the bacterial cell strain E.coli BL21(DE3). Our studies demonstrated the addition of calcium addition in the media does not affect growth but increases protein expression, while reducing the pH from 7 to 4 through the addition of 10mM EGTA in LB media inhibits both. Additionally, we report studies on the design, expression, and purification of fluorescent mCherry variants and their differences in their optical properties, including: extinction coefficients , quantum yields and pKa values. Also, we report progress in the crystallization of two GFP calcium sensors: G1 and D1, using 13 and15% PEG 4000 and 3350 respectively in 50mM HEPES buffer (pH 6.8-7.0) in an effort to optimize crystallization.
DOI
https://doi.org/10.57709/1229028
Recommended Citation
Delgado, Malcom Arturo, "Biochemical Study of Engineered Fluorescent Proteins as Calcium Sensors and the Effect of Calcium and PH in Cell Reproduction and Protein Expression." Thesis, Georgia State University, 2009.
doi: https://doi.org/10.57709/1229028